Culture medium and a method for detection of parasites

Abstract

This invention relates to a culture medium, a kit containing the culture medium and to a method for detection of a parasite such as Dientamoeba fragilis and/or another parasite. The culture medium of the invention is bi-phasic and includes a solid phase containing an egg slope or agar slope; and a liquid phase including a serum and a peptone.

1. A bi-phasic culture medium that supports the growth of protozoa, comprising:
 a solid phase containing an egg slope or agar slope; and 
 a live  Escherichia coli -free liquid phase consisting of:
 a serum, 
 a peptone, 
 a phosphate buffered saline; and 
 optionally an antibiotic or antibiotics, 
 
 wherein the bi-phasic medium supports the growth of protozoa in the presence of only rice starch. 
 
     
     
       2. The medium of  claim 1 , wherein the solid phase is an egg slope. 
     
     
       3. The medium of  claim 1 , wherein the serum is horse serum or rabbit serum. 
     
     
       4. The medium of  claim 1 , wherein the peptone is peptone or bactopeptone. 
     
     
       5. The medium of  claim 1 , wherein pH of the phosphate buffered saline is from about 6.8 to about 7.8. 
     
     
       6. The medium of  claim 1 , wherein the liquid phase is up to about 98 vol % phosphate buffered saline. 
     
     
       7. The medium of  claim 1 , wherein the liquid phase contains about 1 to about 15 vol % of serum. 
     
     
       8. The medium of  claim 1 , wherein the peptone is about 1 to about 40 w/w % bactopeptone solution. 
     
     
       9. The medium of  claim 1 , wherein the liquid phase contains about 1 to about 15 vol % of the peptone. 
     
     
       10. The medium of  claim 1 , wherein the liquid phase includes an antibiotic. 
     
     
       11. The medium of  claim 10 , wherein the antibiotic is selected from the group consisting of erythromycin, penicillin, streptomycin, clindamycin, cephalexin, vancomycin and rifampicin. 
     
     
       12. A kit, comprising:
 a container containing the medium of  claim 1 ; and 
 a container containing rice starch. 
 
     
     
       13. The kit of  claim 12  which is contained in a compartmentalized specimen bag. 
     
     
       14. The kit of  claim 12 , further including a utensil for transferring a specimen into the container containing the medium. 
     
     
       15. The kit of  claim 12  including an additional container for containing a specimen. 
     
     
       16. The kit of  claim 12 , wherein the container containing rice starch is a sachet. 
     
     
       17. A method of detecting the presence of protozoa in a specimen, the method including:
 adding to the medium of  claim 1 , the specimen, rice starch and where necessary, an antibiotic; 
 allowing the medium to incubate for a time period so as to cultivate protozoa; and 
 examining at least a portion of the incubated medium to detect the presence of protozoa. 
 
     
     
       18. A method of detecting protozoa in faecal matter, comprising:
 adding to the medium of  claim 1  faecal matter, rice starch and where necessary, an antibiotic; 
 allowing the medium to incubate for a time period so as to cultivate intestinal protozoa; and 
 examining at least a portion of the incubated medium to detect the presence of the protozoa. 
 
     
     
       19. The method of  17 , wherein the protozoa detected is one or more of  Dientamoeba fragilis, Blastocystis hominis, E. histolytica/dispar, Entamoeba  or  Iodamoeba, Iodamoeba butschlii, Endolimax nana, Entamoeba coli , or  Entamoeba hartmanni . 
     
     
       20. The method of  claim 19 , wherein the protozoa detected is  Dientamoeba fragilis . 
     
     
       21. The method of  claim 17 , wherein the medium is incubated for a period of up to about 4 days. 
     
     
       22. The method of  claim 17 , wherein the medium is incubated for up to about 48 hours. 
     
     
       23. The method of  claim 17 , wherein additional antibiotic and/or rice starch are added after about 24 hours of incubation. 
     
     
       24. The method of  claim 17 , wherein the medium is incubated at a temperature in the range of 36° C. to 38° C. 
     
     
       25. The method of  claim 17 , wherein the portion of the incubated medium is examined microscopically. 
     
     
       26. The method of  claim 17 , wherein the antibiotic is erythromycin. 
     
     
       27. The method of  claim 17 , wherein the portion is or includes sediment. 
     
     
       28. The method of  claim 18 , wherein the protozoa detected is one or more of  Dientamoeba fragilis, Blastocystis hominis, E. histolytica/dispar, Entamoeba  or  Iodamoeba, Iodamoeba butschlii, Endolimax nana, Entamoeba coli , or  Entamoeba hartmanni . 
     
     
       29. The method of  claim 18 , wherein the medium is incubated for a period of up to about 4 days. 
     
     
       30. The method of  claim 18 , wherein additional antibiotic and/or rice starch are added after about 24 hours of incubation. 
     
     
       31. The method of  claim 18 , wherein the medium is incubated at a temperature in the range of 36° C. to 38° C. 
     
     
       32. The method of  claim 18 , wherein the portion of the incubated medium is examined microscopically. 
     
     
       33. The method of  claim 18 , wherein the antibiotic is erythromycin. 
     
     
       34. The method of  claim 18 , wherein the portion is or includes sediment.